MUC1-C influences cell survival in lung adenocarcinoma Calu-3 cells after SARS-CoV-2 infection

Maharjan, Sony; Park, Jeong-A; Kim, Dongbum; Lee, Younghee; Kwon, Hyung‐Joo

doi:10.5483/bmbrep.2021.54.8.018

Cited by 18 publications

(16 citation statements)

References 38 publications

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“…To estimate viral titers, we performed plaque assays on Vero E6 cells, as described previously ( Kim et al, 2021b , 2022a ; Park et al, 2021a ). In brief, Vero E6 cells (7 × 10 5 cells/well) were cultured in 6-well plates (Corning Inc., Corning, NY, United States) for 12 h. Cells were then washed with phosphate-buffered saline (PBS) and infected with 10-fold serial dilutions of virus culture supernatants.…”

Section: Methodsmentioning

confidence: 99%

Differential effect of SARS-CoV-2 infection on stress granule formation in Vero and Calu-3 cells

et al. 2022

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Stress granule formation is induced by numerous environmental stressors, including sodium arsenite treatment and viral infection. Accordingly, stress granules can inhibit viral propagation and function as part of the antiviral host response to numerous viral infections. Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) antagonizes stress granule formation, in part, via interaction between SARS-CoV-2 nucleocapsid (N) protein and Ras-GTPase-activating SH3-domain-binding protein 1 (G3BP1). However, it is unclear whether there are differential effects in different cell types. In this study, we assessed interaction between the N protein of SARS-CoV-2 S clade and G3BP1/2 in Vero and Calu-3 cells and investigated the effect of various SARS-CoV-2 strains on sodium arsenite-induced stress granule formation. Our data show that SARS-CoV-2 S clade N protein interacts with both G3BP1 and G3BP2 more strongly in Calu-3 vs. Vero cells. Consistent with this observation, infection with SARS-CoV-2 S clade induces stress granule formation in Vero but not in Calu-3 cells. However, infection with SARS-CoV-2 S clade, as well as other SARS-CoV-2 variants, inhibits sodium arsenite-induced stress granule formation in both cell lines. Taken together, our results show differential effects of SARS-CoV-2 infection on stress granule formation that is dependent on host cell type, rather than virus strain type.

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Section: Methodsmentioning

confidence: 99%

Differential effect of SARS-CoV-2 infection on stress granule formation in Vero and Calu-3 cells

et al. 2022

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“…Cell culture supernatants were harvested and centrifuged for 10 min at 3,000 rpm to remove cell debris. The cleared supernatants were collected and then virus titers were quantified by plaque assay as described previously ( Kandeel et al, 2020 ; Kim et al, 2021 ). The virus containing media was stored at −70 ° C. MERS-CoV and SARS-CoV-2 amplification and cell culture procedures were performed according to biosafety level 3 (BSL-3) conditions in the Hallym Clinical and Translational Science Institute in accordance with the recommendations of the Institutional Biosafety Committee of Hallym University.…”

Section: Methodsmentioning

confidence: 99%

Production of SARS-CoV-2 N Protein-Specific Monoclonal Antibody and Its Application in an ELISA-Based Detection System and Targeting the Interaction Between the Spike C-Terminal Domain and N Protein

Kim

Baek

et al. 2021

Front. Microbiol.

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SARS-CoV-2 infections continue to spread quickly by human-to-human transmission around the world. Therefore, developing methods to rapidly detect SARS-CoV-2 with high sensitivity are still urgently needed. We produced a monoclonal antibody that specifically detects the N protein of SARS-CoV-2 and recognizes N protein in cell lysates of SARS-CoV-2–infected Vero cells but not in cell lysates of MERS-CoV- or HCoV-OC43-infected Vero cells. This antibody recognized N protein in SARS-CoV-2 clades S, GR, and GH and recognized N protein in all the SARS-CoV-2 clades to ∼300 pfu. Previously, we reported that the coronavirus N protein interacts with the C-terminal domain of the spike protein (Spike CD). In this study, we developed an ELISA-based “bait and prey” system to confirm the interaction between SARS-CoV-2 Spike CD and N protein using recombinant fusion proteins. Furthermore, this system can be modified to quantitatively detect SARS-CoV-2 in culture media of infected cells by monitoring the interaction between the recombinant Spike CD fusion protein and the viral N protein, which is captured by the N protein–specific antibody. Therefore, we conclude that our N protein–specific monoclonal antibody and our ELISA-based bait and prey system could be used to diagnose SARS-CoV-2 infections.

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“…The peptide was conjugated with nine D-arginine residues at the N-terminus (R-Spike CD HCoV-OC43) to penetrate host cells. A negative control cell-penetrating peptide (R-CP-1) was synthesized by Anygen Co., Ltd. (Gwang Ju, South Korea) to contain nine D-arginine residues (NH 2 -d-RRRRRRRRR-AQARRKNYGQLDIFP-COOH) ( 28 ).…”

Section: Methodsmentioning

confidence: 99%

Targeting the Interaction Between Spike Protein and Nucleocapsid Protein for Suppression and Detection of Human Coronavirus OC43

Kim

Kang

et al. 2022

Front. Immunol.

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Human coronavirus OC43 (HCoV-OC43) is the coronavirus most associated with “common colds”, infections of the upper respiratory tract. Previously, we reported that direct interactions of nucleocapsid (N) protein and C-terminal domain of Spike protein (Spike CD) are essential for replication of SARS-CoV-2 and MERS-CoV. Thus, we developed a novel ELISA-based strategy targeting these specific interactions to detect SARS-CoV-2 and MERS-CoV. Here, we investigated whether the same principles apply to HCoV-OC43. We discovered that the S protein of HCoV-OC43 interacts with N protein and that cell penetrating Spike CD peptide inhibits virus protein expression and replication of HCoV-OC43. The interaction between HCoV-OC43 S and N proteins were recapitulated with a recombinant HCoV-OC43 Spike CD fusion protein and a recombinant HCoV-OC43 N fusion protein in vitro. By producing an anti-HCoV-OC43 N protein-specific monoclonal antibody, we established a virus detection system based on the interaction between recombinant Spike CD and N protein of HCoV-OC43. We suggest that the interaction between Spike CD and N protein is conserved in coronaviruses and therefore could be a target for therapeutics against both novel coronavirus and its variants.

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MUC1-C influences cell survival in lung adenocarcinoma Calu-3 cells after SARS-CoV-2 infection

Cited by 18 publications

References 38 publications

Differential effect of SARS-CoV-2 infection on stress granule formation in Vero and Calu-3 cells

Differential effect of SARS-CoV-2 infection on stress granule formation in Vero and Calu-3 cells

Production of SARS-CoV-2 N Protein-Specific Monoclonal Antibody and Its Application in an ELISA-Based Detection System and Targeting the Interaction Between the Spike C-Terminal Domain and N Protein

Targeting the Interaction Between Spike Protein and Nucleocapsid Protein for Suppression and Detection of Human Coronavirus OC43

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